Sutipa TanapongpipatPeerada PromdonkoyToru WatanabeWitoon TirasophonNiran RoongsawangYasunori ChibaLily EurwilaichitrThailand National Center for Genetic Engineering and BiotechnologyNational Institute of Advanced Industrial Science and TechnologyMahidol University2018-06-112018-06-112012-09-01FEMS Microbiology Letters. Vol.334, No.2 (2012), 127-13415746968037810972-s2.0-84865415923https://repository.li.mahidol.ac.th/handle/20.500.14594/13622This study describes Pichia thermomethanolica BCC16875, a new methylotrophic yeast host for heterologous expression. Both methanol-inducible alcohol oxidase (AOX1) and constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoters from Pichia pastoris were shown to drive efficient gene expression in this host. Recombinant phytase and xylanase were expressed from both promoters as secreted proteins, with the former showing different patterns of N-glycosylation dependent on the promoter used and culture medium. In addition, growth temperature also had an effect on N-glycan modification of cell wall mannoproteins. The major glycoprotein oligosaccharide species produced from P. thermomethanolica BCC16875 is Man 8-12 GlcNAc 2 , which is similar to that from other methylotrophs. Moreover, mannosylphosphate and α-1,6- and α-1,2-linked mannose modifications of heterologous secreted protein were also detected. The attainably high level of protein production in complement to distinctive thermotolerance rarely found in other industrial yeasts makes this microorganism an attractive host for large-scale fermentation. © 2012 Federation of European Microbiological Societies.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyImmunology and MicrobiologyArticleSCOPUS10.1111/j.1574-6968.2012.02628.x